Actinomycin D induces apoptosis and inhibits growth of pancreatic cancer cells

Kleeff, Jörg; Kornmann, Marko; Sawhney, Harneet; Korc, Murray

doi:10.1002/(sici)1097-0215(20000501)86:3<399::aid-ijc15>3.0.co;2-g

Cited by 119 publications

(81 citation statements)

References 45 publications

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“…Prolonged exposure to AD was accompanied by intensification of the DAPI signal, suggestive of chromatin condensation. AD is known to induce apoptosis (49). However apoptosis is associated with cell shrinkage, whereas we found a mild but significant increase in nuclear size.…”

Section: Discussioncontrasting

confidence: 48%

High content image cytometry in the context of subnuclear organization

et al. 2009

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The organization of proteins in space and time is essential to their function. To accurately quantify subcellular protein characteristics in a population of cells with regard for the stochasticity of events in a natural context, there is a fast-growing need for image-based cytometry. Simultaneously, the massive amount of data that is generated by image-cytometric analyses, calls for tools that enable pattern recognition and automated classification. In this article, we present a general approach for multivariate phenotypic profiling of individual cell nuclei and quantification of subnuclear spots using automated fluorescence mosaic microscopy, optimized image processing tools, and supervised classification. We demonstrate the efficiency of our analysis by determination of differential DNA damage repair patterns in response to genotoxic stress and radiation, and we show the potential of data mining in pinpointing specific phenotypes after transient transfection. The presented approach allowed for systematic analysis of subnuclear features in large image data sets and accurate classification of phenotypes at the level of the single cell. Consequently, this type of nuclear fingerprinting shows potential for high-throughput applications, such as functional protein assays or drug compound screening. ' 2009 International Society for Advancement of Cytometry

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Section: Discussioncontrasting

confidence: 48%

High content image cytometry in the context of subnuclear organization

et al. 2009

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“…For induction experiments, cells were seeded in 10 cm cell culture plates in 10% FCS growth medium and allowed to attach for 12 h. Growth medium was replaced by serumreduced medium (1% FCS), and supplemented with recombinant TGF-b1 (500 pM), BMP2 (100 ng ml À1 ), FGF2 (10 ng ml À1 ), Shh (500 ng ml À1 ), Ihh (500 ng ml À1 ) (R&D Systems GmbH) and TNF-a (100 ng ml À1 ) (Promega Biosciences Inc., Mannheim, Germany) for 48 h. The doses were determined to ensure the efficacy and absent toxicity of each factor (Nakamura et al, 1997;Kleeff et al, 2000;Li et al, 2003;Kayed et al, 2004;Guo et al, 2006). Afterwards, cell culture supernatants, cell lysates, and mRNA were isolated as described.…”

Section: Cell Culturementioning

confidence: 99%

Regulation and functional role of the Runt-related transcription factor-2 in pancreatic cancer

et al. 2007

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Recent evidence suggests that Runt-related transcription factors play a role in different human tumours. In the present study, the localisation of the Runt-related transcription factor-2 (Runx2), its transcriptional activity, as well as its regulation of expression was analysed in human pancreatic ductal adenocarcinoma (PDAC). Quantitative real-time PCR and immunohistochemistry were used for Runx2 expression and localisation analysis. Runt-related transcription factor-2 expression was silenced using specific siRNA oligonucleotides in pancreatic cancer cells (Panc-1) and immortalised pancreatic stellate cells (IPSCs). Overexpression of Runx2 was achieved using a full-length expression vector. TGF-b1, BMP2, and other cytokines were assessed for their potential to regulate Runx2 expression. There was a 6.1-fold increase in median Runx2 mRNA levels in PDAC tissues compared to normal pancreatic tissues (Po0.0001). Runt-related transcription factor-2 was localised in pancreatic cancer cells, tubular complexes, and PanIN lesions of PDAC tissues as well as in tumour-associated fibroblasts/stellate cells. Coculture of IPSCs and Panc-1 cells, as well as treatment with TGF-b1 and BMP2, led to increased Runx2 expression in Panc-1 cells. Runt-related transcription factor-2 overexpression was associated with decreased MMP1 release as well as decreased growth and invasion of Panc-1 cells. These effects were reversed by Runx2 silencing. In conclusion, Runx2 is overexpressed in PDAC, where it is regulated by certain cytokines such as TGF-b1 and BMP2 in an auto-and paracrine manner. In addition, Runx2 has the potential to regulate the transcription of extracellular matrix modulators such as SPARC and MMP1, thereby influencing the tumour microenvironment.

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“…Apoptosis was induced by overnight treatment with 2 mg/ml actinomycin D (Sigma) (Kleeff et al, 2000). We also used a 24 h treatment with 10 ng/ml TNF-a (Sigma) (Muenchen et al, 2000) or 8 Gy X-ray irradiation (Kyprianou et al, 1997), 48 h before apoptosis detection.…”

Section: Apoptosismentioning

confidence: 99%

Dual activities of galectin-3 in human prostate cancer: tumor suppression of nuclear galectin-3 vs tumor promotion of cytoplasmic galectin-3

et al. 2004

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Galectin-3, a multifunctional lectin, is involved during cancer progression. Previous observations showed that both cytosolic expression and nuclear exclusion of galectin-3 in human prostate cancer cells were associated to progression of the disease. In this study, we examined the biological roles of galectin-3 when expressed either in the nucleus or in the cytosol. LNCaP, a galectin-3-negative human prostate cancer cell line, was used to generate transfectants expressing galectin-3 either in the nucleus or in the cytosol. No changes in cell morphology, proliferation, attachment to laminin-1 or androgen dependency were observed. Cytoplasmic galectin-3 induced significantly increased Matrigel invasion, anchorage-independent growth and in vivo tumor growth and angiogenesis, and decreased inducible apoptosis. Surprisingly, nuclear galectin-3 affected these parameters in an opposite fashion with an overall antitumoral activity. Thus, our study demonstrates that galectin-3 exerts opposite biological activities according to its cellular localization: nuclear galectin-3 plays antitumor functions and cytoplasmic galectin-3 promotes tumor progression.

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Actinomycin D induces apoptosis and inhibits growth of pancreatic cancer cells

Cited by 119 publications

References 45 publications

High content image cytometry in the context of subnuclear organization

High content image cytometry in the context of subnuclear organization

Regulation and functional role of the Runt-related transcription factor-2 in pancreatic cancer

Dual activities of galectin-3 in human prostate cancer: tumor suppression of nuclear galectin-3 vs tumor promotion of cytoplasmic galectin-3

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